Cordier Clément, Haustrate Aurélien, Prevarskaya Natalia, Lehen'kyi V'yacheslav
Department of Biology, Laboratory of Cell Physiology, INSERM U1003, Laboratory of Excellence Ion Channel Science and Therapeutics, Faculty of Science and Technologies, University of Lille, Villeneuve d'Ascq, France.
Front Genet. 2023 Jul 27;14:1215645. doi: 10.3389/fgene.2023.1215645. eCollection 2023.
Transient receptor potential vanilloid subfamily member 6 (TRPV6), a highly calcium-selective channel, has been shown to play a significant role in calcium homeostasis and to participate both and in growth, cell survival, and drug resistance of prostate cancer. Its role and the corresponding calcium-dependent pathways were mainly studied in hormone-dependent human prostate cancer cell lines, often used as a model of early-stage prostate cancers. The goal of the present study was to describe the TRPV6-specific phenotype and signaling pathways it is involved in, using castration-resistant prostate cancer cell lines. RNA sequencing (RNA-seq) was used to study the gene expression impacted by TRPV6 using PC3M PC3M and its derivative PC3M-luc-C6 cell line in its native and TRPV6 overexpressed form. In addition to the whole-cell RNA sequencing, immunoblotting, quantitative PCR, and calcium imaging were used to validate gene status and functional consequences, in both and TRPV6 overexpression cell lines. status was validated using both immunoblotting and quantitative PCR, and the functional consequences of either gene deletion or TRPV6 overexpression were shown using calcium imaging. RNA-seq analysis demonstrated that the calcium channel TRPV6, being a crucial player of calcium signaling, significantly impacts the expression of genes involved in cancer progression, such as cell cycle regulation, chemotaxis, migration, invasion, apoptosis, ferroptosis as well as drug resistance, and extracellular matrix (ECM) re-organization. Our data suggest that the gene is involved in and regulates multiple pathways related to tumor progression and drug resistance in castration-resistant prostate cancer cells.
瞬时受体电位香草酸亚家族成员6（TRPV6）是一种高度钙选择性通道，已被证明在钙稳态中起重要作用，并参与前列腺癌的生长、细胞存活和耐药性。其作用及相应的钙依赖途径主要在激素依赖的人前列腺癌细胞系中进行研究，这些细胞系常被用作早期前列腺癌的模型。本研究的目的是利用去势抵抗性前列腺癌细胞系描述TRPV6特异性表型及其所涉及的信号通路。使用RNA测序（RNA-seq）研究TRPV6对基因表达的影响，采用PC3M及其衍生物PC3M-luc-C6细胞系的天然形式和TRPV6过表达形式。除了全细胞RNA测序外，还使用免疫印迹、定量PCR和钙成像来验证天然细胞系和TRPV6过表达细胞系中的基因状态和功能后果。通过免疫印迹和定量PCR验证基因状态，并使用钙成像显示基因缺失或TRPV6过表达的功能后果。RNA-seq分析表明，钙通道TRPV6作为钙信号的关键参与者，显著影响参与癌症进展的基因表达，如细胞周期调控、趋化性、迁移、侵袭、凋亡、铁死亡以及耐药性和细胞外基质（ECM）重组。我们的数据表明，该基因参与并调节去势抵抗性前列腺癌细胞中与肿瘤进展和耐药性相关的多种途径。
